Apparatus for and method of preparing plant tissue for plant production

ABSTRACT

An apparatus for preparing plant tissue (e.g., somatic embryos, embryogenic tissue, organogenic tissue, vegetative tissue, seeds, etc.) for plant production includes a first station having a first rack system configured to support at least one culture vessel, a second station having an automated member configured to manipulate the at least one culture vessel and a third station having a second rack system configured to support the at least one culture vessel after being manipulated by the automated member. The second station can be selectively adjusted to perform more than one operation required in the plant development. According to exemplary embodiments, the apparatus may include more than one second station (e.g., operational stations, etc.).

CROSS REFERENCE TO RELATED PATENT APPLICATIONS

The present application claims the benefit under 35 U.S.C. 119(e) ofU.S. Provisional Application No. 60/960,282, having a filing date ofSep. 24, 2007, titled “APPARATUS FOR AND METHOD OF PREPARING PLANTTISSUE FOR PLANT PRODUCTION,” the complete disclosure of which is herebyincorporated by reference.

BACKGROUND

The present disclosure relates generally to an apparatus for and amethod of preparing plant tissue (e.g., somatic embryos, embryogenictissue, organogenic tissue, vegetative tissue, seeds, etc.) for plantproduction. More particularly, the present disclosure relates to an atleast partially automated apparatus for and a method of preparing planttissue for plant production.

The process of preparing plant tissue, such as somatic embryos, forplant production (e.g., in the case of conifer treatment, etc.)generally includes the following steps: 1) cone collection and storage;2) somatic embryogenic initiation on an initiation medium; 3)maintenance of embryogenic tissue on a maintenance medium; 4) cryogenicstorage of embryogenic tissue and subsequent cryoretrieval; 5) growth ofembryogenic tissue; 6) development of somatic embryos on an embryodevelopment medium; 7) harvesting of embryos; 8) conditioning ofharvested embryos; and 9) germination.

Plating, inspecting, treating, collecting and/or storing of plantembryos prior to germination are key operations in many steps of theplant production process. The activities necessary for performing theseoperations, however, are usually performed by hand. For example,individual embryos are typically transferred to and from various mediaand vessels and must be plated onto media, one by one using forceps andoften with the guidance of a dissecting microscope.

Such methods are burdensome, time-consuming, costly, and susceptible tocontamination. Not only that, but only a limited number of embryos canbe plated, inspected, treated and/or collected by a single person duringa given period of time. Accordingly, any attempt to increase the numberof embryos that can be produced and subsequently conditioned forgermination necessarily requires an increase in manpower, which itselfcan be costly and often impractical.

There have been attempts to automate certain steps in the production ofplant embryos. For example, U.S. Publication No. 2006/0260015, titled“Somatic Embryogenesis and Embryo Harvesting and Method and Apparatusfor Preparing Plant Embryos for Plant Production” and assigned toArborGen, LLC, the entire disclosure of which is hereby incorporated byreference, discloses an at least partially automated apparatus forwashing and harvesting plant embryos. There have also been attempts toautomate the inspection of plant embryos. However, prior attempts toautomate steps within the production process require a separate piece ofequipment to perform the particular step to be automated. Since theproduction of plant embryos involves multiple steps, requiring aseparate piece of equipment for each step would be costly.

Accordingly, there is a need in the agricultural industry and, inparticular, the forestry sciences, for an apparatus for and a method ofreducing human intervention within the plant production process.Reducing human intervention may reduce the likelihood of contaminationand/or may provide for a more efficient system (e.g., one that can bereadily scaled-up for commercial purposes, etc.). There is also a needfor an apparatus that is capable of performing more than one step withinthe production process so that the amount of equipment needed to providefor an at least partially automated process (and the cost associatedtherewith) can be reduced.

SUMMARY

One exemplary embodiment relates to an apparatus for preparing planttissue for plant production. The apparatus comprises a first stationincluding a first rack system configured to support at least one culturevessel, a second station including an automated member configured tomanipulate the at least one culture vessel and a third station having asecond rack system configured to support the at least one culture vesselafter being manipulated by the automated member.

Another exemplary embodiment relates to an apparatus for preparing planttissue contained within a culture vessel for plant production. Theapparatus comprises a member configured to move the culture vesselbetween an open position and a closed position, a delivery systemconfigured to dispense a substance to the culture vessel when in theopen position, an imaging system configured to capture an image of theplant tissue and a controller coupled to the member, the delivery systemand the imaging system for automating the operation of each.

Another exemplary embodiment relates to a method of preparing planttissue for plant production. The method comprises the steps oftransporting a first culture vessel from a first station to a secondstation, manipulating the first culture vessel between a closed positionand an open position, supplying at least one substance to the firstculture vessel and controlling with a controller the steps oftransporting, manipulating and supplying.

Another exemplary embodiment relates to a method of preparing planttissue for plant production. The method comprises the steps oftransporting a first culture vessel from a first station to a secondstation, performing a first operational step on the first culture vesselat the second station, controlling with a controller the steps oftransporting the first culture vessel and performing the firstoperational step and reconfiguring at least one of the second stationand the controller so that a second operational step can be performed onthe first culture vessel at the second station. The second operationalstep is different than the first operational step.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a perspective view of a plant tissue production apparatusshown according to an exemplary embodiment.

FIG. 2 is a front elevation view of the plant tissue productionapparatus of FIG. 1.

FIG. 3 is a top plan view of the plant tissue production apparatus ofFIG. 1.

FIG. 4 is a side elevation view of the plant tissue production apparatusof FIG. 1.

FIG. 5 is another perspective view of the plant tissue productionapparatus of FIG. 1.

FIG. 6 is a perspective view of a first station of the plant tissueproduction apparatus shown according to an exemplary embodiment.

FIG. 7 is a perspective view of a third station of the plant tissueproduction apparatus shown according to an exemplary embodiment.

FIG. 8 is a perspective view of a second station of the plant tissueproduction apparatus shown according to an exemplary embodiment.

FIG. 9 is another perspective view of the second station of FIG. 8.

FIG. 10 is another perspective view of the second station of FIG. 8.

FIGS. 11A through 11K are schematics illustrating a transportingsequence of operation according to an exemplary embodiment.

FIGS. 12A through 12E are schematics illustrating a dispensing sequenceof operation according to an exemplary embodiment.

FIG. 13 is a perspective view of a culture vessel for use with the planttissue production apparatus of FIG. 1 shown according to an exemplaryembodiment.

FIG. 14 is a side elevation view of the culture vessel of FIG. 13.

FIG. 15 is a perspective view of a top portion of the culture vessel ofFIG. 13.

FIG. 16 is a perspective view of a bottom portion of the culture vesselof FIG. 13.

FIG. 17 is another perspective view of the plant tissue productionapparatus of FIG. 1 showing a reservoir according to an exemplaryembodiment.

DETAILED DESCRIPTION

Referring generally to the FIGURES, an apparatus is provided that isuseful in preparing plant tissue (e.g., somatic embryos, embryogenictissue, organogenic tissue, vegetative tissue, seeds, etc.) forproduction (e.g., large-scale or commercial production, etc.). Theapparatus, referred to broadly herein as a plant tissue productionapparatus, is a multi-functional and at least partially controlled orautomated (e.g., semi-automated, fully-automated, etc.) machine capableof performing operations required in the development and/or productionof plant tissue. The plant tissue production apparatus is configured tobe adjustable (e.g., configurable, reconfigurable, programmable,reprogrammable, etc.) so that it can perform more than one of theoperations required at the various steps in the plant tissue productionprocess.

For example, the plant tissue production apparatus may be selectivelyadjusted to perform one or more of the following operations: 1) mediaplating; 2) tissue plating; 3) process and/or plant tissue inspection;4) quality assessment; and 5) sorting. For purposes of the presentdisclosure, media plating refers to the transferring of a base media(e.g., an initiation media, a maintenance media, a development media,etc.) to a vessel; tissue plating refers to the transferring of a planttissue (e.g., a suspension culture, a gelled culture, etc.) to the basemedia; process and/or plant tissue inspection refers to obtaining animage or other characteristic of the process and/or plant tissue duringdevelopment; quality assessment refers to using the image or othercharacteristic of the plant tissue to determine its quality based uponpredetermined parameters; and sorting refers to the collection planttissue either individually or en masse.

According to an exemplary embodiment, the plant tissue productionapparatus is capable of performing all of the above-mentioned operationsdue to its adjustability (e.g., modularity, flexibility, etc.). Sincethe time between steps (e.g., lag time, development period, conditioningperiod, etc.) is typically a sizable time period, providing a machinethat performs multiple steps reduces the likelihood that the machinewill stay idle or unused between the steps. The plant tissue productionapparatus may also be configured to provide developed or partiallydeveloped plant tissue to another apparatus or module, such as an embryoharvesting system of the type disclosed in U.S. Publication No.2006/0260015 (referenced above) or any other apparatus for which it maybe desirable to combine with the plant tissue production apparatus.

Providing an at least partially automated apparatus capable ofperforming more than one operation by being adjustable mayadvantageously reduce the number of machines required to automate theproduction of plant tissue thereby reducing equipment costs. Also,providing an at least partially automated apparatus capable ofperforming an operation required for the production of plant tissue mayadvantageously improve the efficiency and plant tissue production ratesrelative to hand-operated systems. For example, providing an at leastpartially automated apparatus capable of performing an operationrequired for the production of plant tissue that would otherwise need tobe done manually may improve the quality of the plant tissue, reduce theamount of time needed to produce the plant tissue, reducing the amountof contaminated plant tissues, or increase the overall yield of planttissue. Such efficiency may be realized in any phase or operationperformed in the development process (e.g., plating, inspecting,harvesting, etc.).

Referring to FIGS. 1-4 in particular, a plant tissue productionapparatus 10 is shown according to an exemplary embodiment. Apparatus 10generally includes a first or input station 12, a second or operationalstation 14 and a third or output station 16. Apparatus 10 also includesa transport system 18 configured to selectively transport (e.g.,deliver, move, etc.) one or more receptacles (e.g., containers, trays,plates, Petrie dishes, etc.), shown as incubation or culture vessels 20,between input station 12, operational station 14 and output station 16.It should be noted that the outset that apparatus 10 may include anynumber of operational stations or modules between the input and outputstations. The addition of more than one operational station or modulewithin the system may allow for more flexibility and/or greaterthroughput by providing multiple (e.g., different, etc.) operationssimultaneously.

Referring to FIG. 5, input station 12 is shown according to an exemplaryembodiment. Input station 12 includes a first rack system 40 configuredto support a plurality of culture vessels 20 in an arranged (e.g.,organized, etc.) manner before being presented to operational station14. According to the embodiment illustrated, culture vessels 20 arearranged in a substantially vertical direction at first rack system 40.In particular, culture vessels 20 are shown as being stacked with oneculture vessel 20 being directly supported by another culture vessel 20.To facilitate the stacking arrangement of culture vessels 20, first racksystem 40 includes one or more guide structures (e.g., a track system,etc.), shown as rails 42, extending in a substantially verticaldirection. According to the embodiment illustrated, a single stack(e.g., batch, etc.) of culture vessels 20 is provided at input station12. First rack system 40 is configured to hold between approximately tenand approximately fifteen culture vessels 20, but alternatively, may beconfigured to hold more or less culture vessels 20. Culture vessels 20,the configuration of which is detailed below, are preferably all of thesame size or at least have the same size footprint so that first racksystem 40 does not have to be reconfigured, but alternatively may be ofvarying sizes.

According to the embodiment illustrated, rails 42 are positioned alongthree sides of culture vessels 20 stacked at input station 12. Inparticular, rails 42 are shown as being positioned along a first or leftside, a second or right side and a third or rear side of input station12. Such an arrangement allows culture vessels 20 to be removed fromand/or added to input station 12 from a fourth or front side of inputstation 12 while still providing suitable guidance for culture vessels20.

According to an exemplary embodiment, culture vessels 20 are configuredto move (e.g., slide, etc.) in a substantially vertical directionrelative to rails 42 before being transported to another station withinapparatus 10. During such movement, at least a portion of culturevessels 20 are likely to be in direct contact with one or more of rails42. To reduce the amount of friction (e.g., sliding friction, etc.)existing between culture vessels 20 and rails 42, and/or or reduce theamount of wear on culture vessel 20, friction reducing members (e.g.,bearing surfaces, etc.), shown as pads 44, are coupled to rails 42. Pads44 may be formed of any of a variety of known or otherwise suitablematerials including, but not limited to, plastic (e.g., Delrin, etc.).

Referring to FIG. 6, culture vessels 20 arranged at input station 12 aresupported in a horizontal plane by one or more support members (e.g.,fingers, tabs, projections, etc.), shown as carriers 46. Carriers 46 areshown as being provided along the left and right sides of input station12 and are configured to engage a bottom and/or side portion of thelowermost culture vessel 20 provided in the stack. According to theembodiment illustrated, two carriers 46 are provided along each of theleft and right sides of input station 12. Carriers 46 are movablebetween a first or extended position and a second or retracted positionto allow culture vessels 20 to move to another station within apparatus10. In the extended position, carriers 46 support the entire stack ofculture vessels provided at input station 12 by engaging the lowermostculture vessel 20 provided in stack. In the retracted position, carriers46 release the lowermost culture vessel 20 by moving outward fromculture vessels 20 in a substantially horizontal direction. As detailedbelow, when carriers 46 are in the retracted position, another mechanism(e.g., a component of transport system 18, etc.) is provided at inputstation 12 to support the stack of culture vessels 20.

To facilitate the movement of carriers 46 between the extended positionand the retracted position, one or more movement devices (e.g.,actuators, motors, drives, etc.), shown as linear actuators 48, areprovided. According to an exemplary embodiment, a separate linearactuator 48 is associated with each carrier 46. Each linear actuator 48is coupled to a controller (detailed below) so that the sequence ofmovement of culture vessels 20 between stations can be optimized.According to the various alternative embodiments, a single movementdevice may be provided at each side of input station 12 to control themovement of carriers 46 provided along that side.

One or more sensors can be provided at first rack system 40 to monitorthe number of culture vessels 20 stacked within rails 42. The one ormore sensors can be any type of detection sensor known or otherwisesuitable and can be coupled to rails 42. For example, the one or moresensors can be capacitive, ultrasonic, optical or electrical-contactingsensors. If the one or more sensors detected that the number ofincubations vessels 20 are low or depleted, one or more of the followingactions can be taken: 1) the operation being conducted at operationalstation 14 can be halted; 2) additional culture vessels 20 can bedelivered to first rack system 40; or 3) a warning may be generated viaan alarm or display to warn an operator that the number of culturevessels 20 are low or depleted.

The delivery of culture vessels 20 to first rack system 40 may be doneeither manually (e.g., hand delivery, etc.) or via an automated deliveryprocess. Culture vessels 20 may be delivered to first rack system 40individually or as a batch (e.g., set, etc.) of culture vessels 20. Forexample, a batch of culture vessels 20 may be taken from a growth ordevelopment room (or any other area) and brought to apparatus 10. Asindicated above, culture vessels 20 may be loaded from the front side ofinput station 12, or alternatively, may be loaded from the top of inputstation 12.

Referring to FIGS. 5 and 8-10, operational station 14 is shown accordingto an exemplary embodiment. Operational station 14 constitutes the areaon apparatus 10 at which culture vessels 20 and/or the contents thereinare acted upon (e.g., plated, analyzed, inspected, opened, sorted,collected, etc.). Provided at operational station 14 is a dispensingsystem 50, a manipulation system 52 and an imaging system 54. Accordingto an exemplary embodiment, dispensing system 50 and manipulation system52 are supported by one or more positioning devices or robots to providefor an at least partially automated system. According to the variousalternative embodiments, one or more of dispensing system 50 andmanipulation system 52 may be supported at a relatively fixed orstationary structure (e.g., support frame, housing, etc.).

Referring to FIGS. 8 and 9 in particular, dispensing system 50 is shownaccording to an exemplary embodiment. Dispensing system 50 is configuredto dispense a substance into an culture vessel 20 provided atoperational station 14. According to an exemplary embodiment, dispensingsystem 50 is configured to dispense a base media into culture vessel 20during one step in the production process and to dispense a plant tissue(e.g., embryogenic tissue, etc.) into culture vessel 20 during anotherstep in the production process. Dispensing system 50 generally includesa housing (e.g., container, chamber, collection device, etc.), referredto broadly herein as a reservoir 71 (shown in FIG. 17), for holding thesubstance to be dispensed, one or more outlets (e.g., outputs, etc.),shown as nozzles 72, for directionally dispensing the substance, a pump74 for moving the substance from reservoir 71 to nozzles 72, a supportstructure, shown as a robotic arm 76, for supporting and/or movingnozzles 72, and a conduit system 70 for providing fluid communicationbetween reservoir 71, pump 74 and nozzles 72.

Referring to FIG. 17, reservoir 71 can be used to hold or otherwisecontain a substance that will ultimately exit through nozzles 72.Reservoir 71 may be positioned at apparatus 10, or alternatively, may bepositioned at a remote location relative to apparatus 10 and have aconduit system communicating between the reservoir and apparatus 10.According to the embodiment illustrated, reservoir 71 includes a singlecontainer (e.g., bottle, beaker, etc.) defining a receptacle forcontaining a substance to be dispersed from nozzles 72. According to thevarious alternative embodiments, the reservoir may include any number ofcontainers defining receptacles for containing substances to bedispersed from nozzles 72. The multiple containers may contain the samesubstance, or alternatively, may be used to hold different substancesthat can be mixed and/or used independently of the other substances.According to further alternative embodiments, the reservoir may bedivided into separate compartments so that a single reservoir can holddifferent substances simultaneously. Reservoir 71 is a modular systemconfigured to be interchanged with a second reservoir depending upon theparticular operation being conducted at operational station 14 and/orwhen a first reservoir is emptied and/or when a reservoir is no longerneeded within apparatus 10.

Reservoir 71 is shown as being substantially transparent so that anoperator can see through and monitor or observe the substance containedtherein. Reservoir 71 may be formed of any of a number of materialsincluding, but not limited to, a clear polycarbonate, transparent glass,or other type of transparent material. Reservoir 71 can be any shape,such as cylindrical, pyramidal, conical, or cubical. Further, one ormore sensors can be used to monitor the substance level within reservoir71 and to send a signal to the controller (detailed below) that thecontroller may use to cause a particular action to take place (e.g.,stopping of the operation, refilling of reservoir 71, generation of analarm or an informational display, etc.).

As indicated above, dispensing system 50 may be used in plating anembryogenic tissue into culture vessel 20. In certain situations, it isdesirable to “bulk-up” embryogenic tissue before transferring it onto anembryo development media. For example, embryogenic tissue cultures thathave been cryogenically-stored, for instance, are plated onto gelledmedium and incubated for a period of time until there is sufficientgrowth to justify their transfer to a development medium.

The embryogenic tissue may also be “bulked-up” or grown in a liquidversion of the traditional gel medium. Eliminating the plating step mayhelp to streamline the embryo development process and reduce costsassociated with making the gel plates. For this liquid version, liquidsuspension cultures are established by initially dispersing embryogenictissue in liquid media in an appropriately sized flask or bioreactor.Additional liquid suspension medium can be routinely added during theincubation period. Cultures can be monitored until they have grown to amass that is suitable for plating for embryo development.

Embryogenic tissues that have been bulked up from either the traditionalgel or the alternative liquid suspension media can be used to developsomatic embryos. An amount of the bulked up tissue can be transferred toculture vessel 20 and placed onto the surface of embryo developmentmedium. For the liquid version, or in any other application in which itmay be desirable, the flask or bioreactor itself may be coupled to orused as reservoir 71 of dispensing system 50.

Referring to FIG. 9 in particular, nozzles 72 are shown according to anexemplary embodiment. Nozzles 72 constitute the exit for the substancecontained within the reservoir before it is applied in culture vessels20. According to the embodiment illustrated, dispensing system 50 isshown as having a plurality of nozzles 72 supported by robotic arm 76.In particular, nozzles 72 are shown as being spaced apart from eachother in a generally linear manner along robotic arm 76 and aregenerally fixed relative thereto. Nozzles 72 cooperate to span adistance that is substantially equal to at least one dimension ofculture vessel 20 (e.g., the depth of culture vessel 20, etc.) so that asubstance can be equally applied to the entire culture vessel 20 ifdesired.

The number and configuration of nozzles 72 used at operational station14 can be selected depending upon the desired flow rate and/or lay down(e.g., spray, etc.) pattern of the substance. For example, the velocityor pressure of the substance exiting nozzles 72 may be selected suchthat a consistent layer of the substance is applied to culture vessel20. The velocity of the fluid is dependent upon the line pressure andthe design of the nozzles. The velocity or pressure of the substance canbe changed for different types of substances by simply changing thenozzles. It should be noted that substance (e.g., suspension, etc.)being dispersed from nozzles 72 may be a relatively viscous substance(e.g., gel-like, etc.) or a relatively thin substance (e.g., runny,diluted, etc.).

The selection of nozzles 72 also can be based on the desired lay downpattern of dispensing system 50. For example, if the substance beingdispersed from nozzles 72 is a relatively viscous substance, nozzles 72may selected for their ability to lay down relatively continuous rows ofthe substance. Alternatively, if the substance being dispersed fromnozzles 72 is a relatively thin substance (thereby resulting in aspray), a conical spray pattern may be desired in which the sprayimpinging on culture vessel 20 and/or base media contained therein hasan even distribution. Alternatively, the spray pattern may be in a moreannular pattern in which more of the relatively thin substance isdirected toward the center of the spray while there is less substancearound the spray's periphery. One with ordinary skill in the art, oncemade aware of this disclosure, can determine suitable nozzles based onthe desired lay down pattern and the line pressure.

It should be noted that while the embodiment illustrated shows the useof multiple nozzles, according to the various alternative embodiments, asingle nozzle may be used. Such a nozzle may be supported by a fixed ormovable structure depending on the application and may have any of avariety of configurations.

Referring back to FIGS. 8 and 17 in particular, pump 74 is shownaccording to an exemplary embodiment. Pump 74 is coupled betweenreservoir 71 and nozzles 72 and is configured to transfer the substancecontained with reservoir 71 to nozzles 72 for distribution to culturevessel 20. According to an exemplary, pump 74 is a peristaltic pumphaving a pump speed of between approximately 100 and approximately 600revolutions per minute. For example, pump 74 may have a pump speed ofapproximately 270 revolutions per minute. According to the variousalternative embodiments, pump 74 may be selected to have any of avariety of performance capabilities depending on the particularapplication.

Still referring to FIGS. 8 and 17, conduit system 70 is shown accordingto an exemplary embodiment. Conduit system 70 includes suitable plumbing(e.g., tubing, piping, valves, etc.) provided at an input side of pump74 and at an output side of pump 74 for communicating with reservoir 71and nozzles 72 respectively. According to the embodiment illustrated,separate tubing is provided between reservoir 71 and pump 74 and betweenpump 74 and each nozzle 72. According to the various alternativeembodiments, one or more tubes may be coupled to a manifold system influid communication with nozzles 72. Such a manifold system may besupported at robotic arm 76. According to an exemplary embodiment, thetubing used between reservoir 71 and the input of pump 74 and betweenthe output of pump 74 and nozzles 72 has an inside diameter betweenapproximately 0.06 inches and approximately 0.31 inches. For example,the tubing between reservoir 71 and the input of pump 74 and between theoutput of pump 74 and nozzle 72 may have an inside diameter ofapproximately 0.12 inches. According to the various alternativeembodiments, tubing of any size may be used depending on the particularapplication.

Referring to FIGS. 5 and 8, robotic arm 76 is shown according anexemplary embodiment. Robotic arm 76 is configured to support nozzles 72in a movable manner relative an culture vessel 20 provided atoperational station 14. According to the embodiment illustrated, roboticarm 76 is a substantially rectangular member extending outward in adirection that is substantially perpendicular to the length of apparatus10 (i.e., robotic arm 76 extends outward in a direction that issubstantially parallel to a Y-axis of apparatus 10). Robotic arm 76 isconfigured to move from side-to-side while remaining in substantiallythe same horizontal plane. To facilitate the movement of robotic arm 76,one or more movement devices (e.g., actuators, motors, etc.) isprovided. In particular, a linear actuator (not shown) is coupled to arear portion of robotic arm 76 to selectively move robotic arm 76 fromside-to-side (i.e., along the width of operational station 14). Thelinear actuator is coupled to a controller (detailed below) so that itsmovement can be at least partially automated. According to the variousalternative embodiments, robotic arm 76 may 3-degrees-of-freedom (i.e.,movement in an X-direction, Y-direction and Z-direction) for movingnozzles 72 in a variety of directions. According to further alternativeembodiments, robotic arm 76 may up to 6-degrees-of-freedom (i.e.,movement in an X-direction, Y-direction and Z-direction, plus rotationalmovement in three axes for complete maneuverability of nozzles 72 or anyother device supported by robotic arm 76 (e.g., embryo manipulators,etc.)).

When operational station 14 of apparatus 10 is not being used fordispensing a substance, robotic arm 76 is moved to a storage position,which may be located at a periphery of operational station 14 (e.g., theleft side, etc.), to avoid the possibility of interfering with anotherstep that may later occur at operational station 14 (e.g., an inspectionof culture vessels 20 using imaging system 54, etc.). Since nozzles 72may continue to dispense (e.g., leak, drip, etc.) a substance whilerobotic arm 76 is in the storage position, a collection device (e.g.,tray, reservoir, etc.), shown in FIG. 5 as a drip plate 59, is providedunder nozzles 72 to collect any substance being discharged. Drip plate59 preferably retains any such substance and/or directs it so that thesubstance does not contaminate and/or damage other components ofapparatus 10.

Referring to FIGS. 9 and 10, manipulation system 52 is shown accordingto an exemplary embodiment. Manipulation system 52 is configured toengage an culture vessel 20 provided at operational station 14 to movethe culture vessel 20 between an opened position and a closed position.Manipulation system 52 includes one or more operating tools coupled to asupport structure, shown as a robotic arm 58, for engaging culturevessels 20. According to the embodiment illustrated, the one or moreoperating tools include a pair of vacuum heads 56 coupled to a vacuumsystem. Relying upon a negative pressure differential generated by thevacuum system, vacuum heads 56 are configured to engage and hold a coverportion of culture vessel 20 away from a base portion of culture vessel20 while culture vessel 20 is undergoing an operation at operationalstation 14. According to the various alternative embodiments, vacuumheads 56 may be replaced with any other tool suitable for moving culturevessels 20 between the opened and closed positions (e.g., a grippingdevice, etc.).

According to the embodiment illustrated, robotic arm 58 is configured tomove in a substantially vertical direction so that vacuum heads 56 canengage and move the cover portion of culture vessel 20 relative to thebase portion. To facilitate the movement of robotic arm 58, one or moremovement devices (e.g., actuators, motors, etc.) is provided. Inparticular, a linear actuator 53 is coupled to a rear portion of roboticarm 58 to selectively move robotic arm 58 in a substantially verticaldirection (i.e., up and down). Linear actuator 53 is coupled to acontroller (detailed below) so that its movement can be at leastpartially automated.

According to the various alternative embodiments, manipulation system 52may also include one or more additional robotic arms having operatingtools for manipulating the contents of an culture vessel 20 provided atoperational station 14 after being opened by robotic arm 58. Theseadditional robotic arms may be configured to selectively perform atleast one of the following operations: 1) place a plant tissue ontoculture vessel 20; and/or 2) collect plant tissue (e.g., somaticembryos, etc.) from culture vessel 20. The operating tools of theseadditional robotic arms may include a gripping device (e.g., a claw,tweezers, etc.), a suction device (e.g., coupled to a vacuum system,etc.), or any other known or otherwise suitable device for performingany one of the operations just mentioned.

Such robotic arms may include an operating head configured to support aplurality of operating tools. One or more of the operating toolssupported at the operating head can be interchangeable so that apparatus10 can be reconfigured (e.g., retooled, etc.) for different steps in theplant embryo production process. The operating tools can be used aloneor in combination with another tool supported at the operating head(e.g., a camera of an imaging system, etc.). The robotic arm and/or theoperating head may have up to 6-degrees-of-freedom (i.e., movement in anX-direction, Y-direction and Z-direction, plus rotational movement inthree axes for complete maneuverability of embryo manipulation oroperating tools). To facilitate the desired movement of the robotic armand/or the operating head, a first linear actuator (i.e., X-directionlinear actuator), a second linear actuator (i.e., Y-direction linearactuator) and a third linear actuator (i.e., Z-direction linearactuator) may be provided along with actuators (e.g., servo motors,etc.) that provide rotational movement.

Each linear actuator may be driven by a separate motor (e.g., a servomotor, etc.) that provides for the linear motion of the robotic armand/or the operating head along its respective axis. Each linearactuator may also include a guide (e.g., rail, shaft, etc.) to providefor its linear movement. The first linear actuator may selectively movethe operating head side-to-side (e.g., right and left, etc.) or alongthe X-axis of apparatus 10. The second linear actuator may selectivelymove the operating head in-and-out (e.g., front to back, etc.) or alongthe Y-axis of apparatus 10. The third linear actuator may selectivelymove the operating head up-and-down or along the Z-axis of apparatus 10.To provide rotational movement for the robotic arm and/or operatinghead, the system may include a combination of polar axes driven by servomotors. The actuators (both linear and rotational) may be operatedseparate from the other actuators or simultaneously with one or more ofthe other actuators.

Referring back to FIG. 5, imaging system 54 is shown according to anexemplary embodiment. Imaging system 54 is configured to obtain imagesof the contents of an culture vessel 20 (e.g., plant tissue, such assomatic embryos, etc.). According to an exemplary embodiment, imagingsystem 54 includes one or more cameras 55 (e.g., digital imagingcameras, charged coupled devices (CCD), etc.) supported above theposition of an culture vessel 20 provided at operational station 14.While only a single camera may be used, use of additional cameras mayimprove the resolution of the inspection. According to the embodimentillustrated, imaging system 54 includes four cameras 55. Each camera 55is provided to obtain an image of approximately one fourth of an culturevessel 20 (e.g., one quadrant, etc.) provided at operational station 14and is fixedly coupled to a support member 57 extending laterally acrossoperational station 14. Cameras 55 may be operatively coupled to adisplay screen, a controller (detailed below) and/or suitable imageprocessing software.

Imaging system 54 may be used to obtain any of a number ofcharacteristics of the plant tissue including, but not limited to, size,shape, development, symmetry, color, quantity, etc. By obtaining imagesof the characteristics, imaging system 54 may be useful in the steps ofplant embryo inspection, embryo quality assessment and/or embryosorting. During inspection, the images of the plant embryos may bestored in a database system so that their development can be trackedthrough several stages of the production process. When its desirable tomake a quality assessment of each plant embryo, the image processingsoftware may be used to make such a determination. The image processingsoftware is able to make a qualitative decision as to whether each plantembryo is to be considered “quality” or “non-quality” based uponpredetermined parameters of one or more characteristics of the plantembryos. Based upon the quality assessment determined by the imageprocessing software, the controller may direct one or more of therobotic arms of manipulation system 52 to act accordingly (e.g., tocollect only those plant embryo determined to be quality, etc.).

In addition to imaging system 54, apparatus 10 may include additionalinspection systems intended to monitor development within culturevessels 20. For example, apparatus 10 may include inspection systemsconfigured to inspect chemical properties within the vessels (e.g., pH,etc.), media properties (e.g., depth, etc.) and/or any other propertiesthat may be desirable to inspect.

Referring to FIG. 7, apparatus 10 further includes output station 16.Output station 16 is shown as having a second rack system 80 configuredto support a plurality of culture vessels 20 in an arranged manner afterpassing through operational station 14. According to an exemplaryembodiment, the configuration of second rack system 80 is substantiallysimilar to the configuration of first rack system 40. As such, culturevessels 20 are arranged (e.g., stacked, etc.) in a substantiallyvertical direction at second rack system 80 with one culture vessel 20being directly supported by another culture vessel 20. Second racksystem 80 also includes one or more guide structures (e.g., tracksystem, etc.), shown as rails 82, extending in a substantially verticaldirection. Like first rack system 40, second rack system 80 isconfigured to hold up to between approximately ten and approximatelyfifteen culture vessels 20, but alternatively, may be configured to holdmore or less culture vessels 20.

According to an exemplary embodiment, culture vessels 20 are configuredto move (e.g., slide, etc.) in a substantially vertical directionrelative to rails 82 when additional culture vessels 20 are added tooutput station 16 from operational station 14. During such movement, atleast a portion of culture vessels 20 are likely to be in direct contactwith one or more of rails 82. To reduce the amount of friction (e.g.,sliding friction, etc.) existing between culture vessels 20 and rails82, and/or or reduce the amount of wear on culture vessel 20, frictionreducing members (e.g., bearing surfaces, etc.), shown as pads 84, arecoupled to rails 82. Pads 84 may be formed of any of a variety of knownor otherwise suitable materials including, but not limited to, plastic(e.g., Delrin, etc.).

Similar to input station 12, culture vessels 20 arranged at outputstation 16 are supported in a horizontal plane by one or more supportmembers (e.g., fingers, tabs, projections, etc.), shown as carriers 86.Carriers 86 are shown as being provided along the left and right sidesof output station 16 and are configured to engage a bottom and/or sideportion of the lowermost culture vessel 20 provided in the stack.According to the embodiment illustrated, two carriers 86 are providedalong each of the left and right sides of output station 16. Carriers 86are movable between a first or extended position and a second orretracted position to allow output station 16 to receive additionalculture vessels 20 from operational station 14. In the extendedposition, carriers 86 support the entire stack of culture vessels 20provided at output station 12 by engaging the lowermost culture vessel20 provided in stack. In the retracted position, carriers 86 release thelowermost culture vessel 20 by moving outward from culture vessels 20 ina substantially horizontal direction (i.e., the Y-axis of apparatus 10).As detailed below, when carriers 86 are in the retracted positionanother mechanism (e.g., a component of transport system 18, etc.) isprovided at output station 12 to support the stack of culture vessels20.

To facilitate the movement of carriers 86 between the extended positionand the retracted position, one or more movement devices (e.g.,actuators, motors, etc.), shown as linear actuators 88, are provided.According to an exemplary embodiment, a separate linear actuator 88 isassociated with each carrier 86. Each linear actuator 88 is coupled to acontroller (detailed below) so that the sequence of movement of culturevessels 20 between stations can be optimized. According to the variousalternative embodiments, a single movement device may be provided alongeach side of input station 12 to control the movement of carriers 86provided along that side.

One or more sensors can be provided at second rack system 80 to monitorthe number of culture vessels 20 stacked within rails 82. The one ormore sensors can be any type of detection sensor known or otherwisesuitable and can be coupled to rails 82. For example, the one or moresensors can be capacitive, ultrasonic, optical or electrical-contactingsensors. If the one or more sensors detected that the number ofincubations vessels 20 are low or depleted, one or more of the followingactions can be taken: 1) the operation being conducted at operationalstation 14 can be halted; 2) culture vessels 20 can be removed fromsecond rack system 80; or 3) a warning may be generated via an alarm ordisplay to warn an operator that the number of culture vessels 20 ishigh or at its limit.

The removal of culture vessels 20 to first rack system 40 may be doneeither manually (e.g., hand delivery, etc.) or via an automated deliveryprocess. Culture vessels 20 may be removed from first rack system 40individually or as a batch (e.g., set, etc.) of culture vessels 20. Forexample, a batch of culture vessels 20 may be removed from second racksystem 80 after undergoing an operation at operational station 14 (e.g.,tissue plating, etc.) and taken to a growth or development room for anextended period of time. Due to the configuration rails 82, culturevessels 20 may be removed from the front of output station 16 and/or thetop. As detailed, above an identification device may be provided onculture vessels 20 that be used to track the progress of the contents ofthe culture vessels and/or may be used by an automated delivery orwarehousing system.

To facilitate the movement of culture vessels 20 between input station12, operational station 14 and output station 16, transport system 18 isprovided. According to an exemplary embodiment, transport system 18 isconfigured to de-stack an culture vessel 20 supported at input station12, move the de-stacked culture vessel 20 from input station 12 tooperational station 14, move the culture vessel 20 provided atoperational station 14 to output station 16 and re-stack the culturevessel 20 coming from operational station 14 at output station 16. In aneffort to avoid disrupting the development of the plant embryos,transport system 18 is configured to move culture vessels 20 in asubstantially horizontal direction between input station 12, operationalstation 14 and output station 16. In a further effort to limit thedisruption of the contents of culture vessel 20 during movement, culturevessels 20 are taken from the bottom of the stack at first rack system40 and returned to the bottom of the stack at second rack system 80.

Referring to FIGS. 11A-11K, transport system 18 is shown according to anexemplary embodiment. Transport system 18 is shown as including a firstvertical movement device (e.g., lift device, linear actuator, telescopiccylinder, etc.), shown as a first actuator 100, a second verticalmovement device (e.g., lift device, linear actuator, telescopiccylinder, etc.), shown as a second actuator 102, a horizontal movementdevice (e.g., linear actuator, telescopic cylinder, conveyor, etc.),shown as an actuator 104, and a support structure 106 for coupling firstactuator 100 and second actuator 102 to an operating portion of thirdactuator 104.

According to an exemplary embodiment, first actuator 100 issubstantially similar to second actuator 102. First actuator 100 andsecond actuator 102 are configured to move in a substantially verticaldirection to remove and/or present an culture vessel 20 from or to inputstation 12, operational station 14 and output station 16. First actuator100 and second actuator 102 include a support member (e.g., platform,engaging device, etc.), shown as a lift plate 108 and 110 respectively,for supporting culture vessel 20.

First actuator 100 and second actuator 102 are also configured to movein a substantially horizontal direction (e.g., side-to-side, etc.).According to an exemplary embodiment, first actuator 100 is configuredto be selectively moved in the horizontal direction between inputstation 12 and operational station 14, while second actuator 102 isconfigured to be selectively moved in the horizontal direction betweenoperational station 14 and output station 16. The positioning of firstactuator 100 relative to second actuator 102 in the horizontal directionremains constant due to support structure 106. First actuator 100 isspaced apart from second actuator 102 along support structure 106 sothat first actuator 100 will be positioned at either input station 12 oroperational station 14 when second actuator 102 is positioned at eitheroperational station 14 or output station 16 respectively. According tothe embodiment illustrated, support structure 106 includes asubstantially linear member extending between a first end configured tosupport first actuator 100 and a second end configured to support secondactuator 102. A bracket (e.g., plate, etc.) couples the linear member tothird actuator 104 using one or more fastening techniques (e.g., one ormore mechanical fasteners, such as bolts, rivets, screws, etc., awelding operation, etc.).

The method of moving an culture vessel 20 between input station 12,operational station 14 and output station 16 is sequentially shown inFIGS. 11A-11K. In FIG. 11A, all of culture vessels 20 are provided atinput station 12, with first actuator 100 provided at input station 12and second actuator 102 provided at operational station 14. In FIG. 11B,first actuator 100 is moved upwards in a vertical direction to engagethe bottom of the lowermost culture vessel 20. Once engaged, carriers 46are moved to the retracted position so that first actuator 100 is nowsupporting the entire stack of culture vessels 20. First actuator 100then moves slightly downwards (e.g., a distance substantially equal tothe depth of one culture vessel 20, etc.) and carriers 46 are moved backto the extended position to support the remaining stack of culturevessels 20. In FIG. 11C, first actuator 100 is moved downward to aretract position while supporting one culture vessel 20. In FIG. 11D,first actuator 100 and second actuator 102 are moved in a horizontaldirection so that first actuator 100 is provided at operational station14 and second actuator 102 is provided at output station 16.

In FIG. 11E, the culture vessel 20 at operational station 14 is engagedand supported by manipulation system 52. First actuator 100 and secondactuator 102 are then moved back in the horizontal direction so thatfirst actuator 100 is once again provided at input station 12 and secondactuator is provided at operational station 14 as shown in FIG. 11F. InFIGS. 11G and 11H, second actuator 102 now supports the first culturevessel 20 taken from input station 12 and first actuator 100 nowsupports a second culture vessel 20 taken from input station 12. InFIGS. 11I-11K, first actuator 100 and second actuator 102 are moved inthe horizontal direction so that first actuator 100 is once againprovided at operational station 14 and second actuator 102 is once againprovided at output station 16. With second actuator 102 at outputstation 16, the first culture vessel 20 taken is re-stacked at outputstation 16 and supported by carriers 86. The process is repeated untilall of the culture vessels 20 pass through operational station 14.

According to the various alternative embodiments, transport system 18may include any of a number of known or otherwise suitable devices orcomponents for moving culture vessels 20 between input station 12,operational station 14 and output station 16. According to anotherexemplary embodiment, transport system 18 may be configured to deliverculture vessels 20 to another operational system after passing throughoperational station 14. For example, transport system 18 may beconfigured to deliver culture vessels 20 to an automated mass harvestingsystem of the type discloses in U.S. Publication No. 2006/0260015(referenced above). For such an embodiment, the mass harvesting systemmay be configured as a module that can be selectively added or removedfrom apparatus 10.

Apparatus 10 further includes a controller 90 for controlling at leastone, and preferably all, of first rack system 40, second rack system 80,transport system 18, dispensing system 50, manipulation system 52 andimaging system 54, either automatically or by operator control.Controller 90 may be connected to and configured to control thecomponents of these systems by one or more wire transmission linesextending between the various devices that it operates and to thesensors which send it information or by a wireless interface.

Controller 90 may comprise a display, one or more microprocessors,memories, input/output lines, a graphical user interface, and/or one ormore operation buttons. Controller 90 can include, for example, a smallProgrammable Logic Controller (PLC) with an operator interface foroperator inputs, operational parameters, error messages, and productionreports. A PLC with more digital inputs and outputs or a PC-basedcomputer can be used for a fully automated system. For example, thecontroller may contain data processing programs in one or moremicroprocessors for processing data related to the sensors and programsfor performing operational commands for controlling, linear actuators48, robotic arm 58, vacuum heads 56, cameras 55 and/or the variousmovement devices of transport system 18. Furthermore, the controller canbe configured to control the output flow from nozzle 72 by controllingpump 74 and/or one or more valves or regulators associated therewith.

Controller 90 can be programmed to make an entire operation automaticfrom the time when a batch of culture vessels 20 are loaded at inputstation 12 to the time when they are removed from output station 16.Alternatively, controller 90 can be programmed to make only portions ofan operation automatic. For example, the operation taking place atoperational station 16 (e.g., plating, inspection, collecting, etc.) canbe automated while the movement of culture vessels 20 between inputstation 12, operational station 14 and output station 16 areoperator-controlled either manually (by hand) or via the controller.Another example can be to have the entire operation automated whileproviding an operator with the option to halt the operation if desired.For example, if the operator wishes to have an culture vessel undergo anadditional inspection operation, the operation can use the controller tohalt the entire inspection operation and repeat the inspectionoperation.

Referring back to FIGS. 1 and 2, controller 90 is shown as including anON/OFF switch 92, an interface 94, and a display 96. Display 96 may bepart of interface 94 or may be a separate component. Interface 94 allowsan operator to input parameters for the particular operation beingperformed. Interface 94 may be a graphical interface, a touch screeninterface, a keyboard or any other known or otherwise suitable userinterface. Display 96 may display operational variables and parametersto the operator. For example, in the case of an embryogenic tissueplating operation, the number of plating operations performed, the speedof pump 74, the flow rate at nozzle 72, any warnings regarding thesensors at first rack system 40 or second rack system 80, etc. can beshown on display 96. Further, if culture vessels 20 include anidentification device, information stored thereon can be shown ondisplay 96 and made available to an operator.

Still referring to FIGS. 1 and 2, and according to an exemplaryembodiment, input station 12, operational station 14, output station 16and transport system 18 of apparatus 10 are provided within a relativelysterile or low contamination environment (e.g., a clean room, etc.).According to the embodiment illustrated, to facilitate a relativelysterile or low contamination environment for the plant embryosthroughout the production process, input station 12, operational station14, output station 16 and transport system 18 of production apparatus 10are located within a relatively sterile enclosure (e.g., HEPA-filteredchamber, etc.), shown as a laminar flow hood 23. According to anexemplary embodiment, laminar flow hood 23 has a width that isapproximately sixty inches, a depth that is approximately twenty inchesand a height that is approximately twenty-eight inches. Providing thecomponents of production apparatus 10 within laminar flow hood 23reduces the likelihood that the plant embryos will be contaminatedduring the production process. In the case of use in a laminar flowhood, it is important to optimize the design and orientation of thecomponents of the apparatus so as to minimize the redirection of air inthe hood.

According to an exemplary embodiment wherein the components of apparatus10 are enclosed within laminar flow hood 23, controller 90 is providedoutside of laminar flow hood 23. Such positioning of controller 90 mayadvantageously allow an operator to access ON/OFF switch 92, interface94, and/or display 96 of controller 90 without breaching the environmentmaintained within laminar flow hood 23 for the plant embryos. Further,such positioning of controller 90 may prolong the useful life ofcontroller 90 by shielding the components of controller 90 (e.g.,electronics, etc.) from an environment (e.g., relatively high humiditylevels, etc.), that may adversely affect their continued operation.

It should be noted that input station 12, operational station 14, outputstation 16 and/or transport system 18 of apparatus 10 can be usedeffectively outside of a relatively sterile or low contaminationenvironment. For example, certain applications may be less sensitive andmay not require a relatively sterile or low contamination environment toachieve the results desired. In such applications, apparatus 10 may beset up in any location that is convenient.

Referring now to FIGS. 13-16, culture vessel 20 is shown according to anexemplary embodiment. Culture vessel 20 provides a structure suitablefor supporting the development of plant tissue throughout various stepsof the production process. For example, culture vessel 20 can be usedfor plating, inspection, quality assessment, growth, harvesting,conditioning, etc. Culture vessel 20 is designed to hold both liquid andsolid substances. For example, culture vessel 20 is designed to hold abase media material, an embryogenic tissue and one or more developedplant embryos. According to an exemplary embodiment, culture vessel 20is sized to support a plurality of plant embryos (e.g., 10, 100, 1000,10,000, etc.) through the development process. According to furtherexemplary embodiments, culture vessel 20 is configured to hold anynumber of plant tissues through the various stages of development.

Culture vessel 20 is shown as a substantially rectangular (e.g., square,etc.) container including a first portion (e.g., lid, closure, top,etc.), shown as a cover portion 22, coupled to a second portion (e.g.,bottom, receptacle, etc.), shown as a base portion 24. Cover portion 22is coupled to base portion 24 such that culture vessel 20 may beselectively moved between a first or closed position (wherein thecontents of culture vessel 20 are substantially concealed) and a secondor open position (wherein the contents of culture vessel 20 can bephysically accessed). According to an exemplary embodiment, culturevessel 20 has a width of approximately nine inches, a length ofapproximately nine inches and a height of approximately three fourths ofan inch. According to the various exemplary embodiments, culture vessel20 may be provided in any of a number of sizes.

It should further be noted that culture vessel 20 may be configured in awide variety of shapes to accommodate varying design criteria, forexample, as a generally rectangular shaped vessel as illustrated in theFIGURES. According to the various alternative embodiments, culturevessel may be configured as other well-known or otherwise suitableshapes having linear surface and/or nonlinear edges and surfaces. Forexample, culture vessel 20 may be a generally circular, octagonal, etc.Further still, culture vessel 20 may be designed for use during only oneor more specific steps in the development process.

Referring to FIG. 15 in particular, cover portion 22 has an end wall 26(e.g., platform, top, top surface, etc.) and a side wall 28 (skirt,peripheral surface, etc.) extending downward therefrom at an orientationthat is angled slightly outward from end wall 26. According to anexemplary embodiment, end wall 26 includes a relatively flat outersurface that facilitates the organization of the vessels relative toeach other (e.g., by stacking of one culture vessel on top of another,etc.). According to the various alternative embodiments, cover portion22 may not include a side wall or may included a side wall configured tosubstantially align with or fit within a corresponding structure on baseportion 24.

Referring to FIG. 16, base portion 24 is shown according to an exemplaryembodiment. Base portion 24 has an end wall 30 (e.g., platform, bottom,bottom surface, etc.) and a side wall 32 extending upward therefrom atan orientation that is angled slightly outward from end wall 30. Sidewall 32 is designed to be at least partially covered (e.g., overlapped,etc.) by side wall 28 of cover portion 22 when culture vessel 20 is inthe closed position. Side wall 32 defines an aperture 34 (e.g., cavity,receptacle, etc.) suitable for receiving substances used in theproduction process including, but not limited to media for plating andtissue cultures. The size and shape of aperture 34 may vary depending ona number of factors, including the size, shape, and quantity of articlesto be provided therein. Aperture 34 may be divided into one or morecompartments (e.g., sections, storage wells, etc.) for separating one ormore plant embryos during the production process.

According to the embodiment illustrated, there is no latching deviceprovided between cover portion 22 and base portion 24 to secure coverportion 22 in the closed position. In such an embodiment, the weight ofcover portion 22, in combination with the overlap existing between sidewall 28 and side wall 32, help retain cover portion 22 in the closedposition. Since culture vessel 20 does not include a latching device,removal of cover portion 22 (either manually or by apparatus 10) may bedone quickly and more efficiently. For example, according to theembodiment illustrated, cover portion 22 can be removed by moving atleast one of cover portion 22 and base portion 24 relative to the otherone of cover portion 22 and base portion 24 in a substantially verticaldirection. According to the various alternative embodiments, culturevessel 20 may include any known or otherwise suitable latching devicefor securing cover portion 22 to base portion 24 in the closed position.

According to an exemplary embodiment, top portion 22 and base portion 24are formed of a relatively transparent material so that objects withinculture vessel 20 can be readily inspected without requiring culturevessel 20 to be moved to the open position. Further, as detailed above,culture vessel 20 is often placed within a growth or development roomfor an extended period of time after an operation is performed byapparatus 10. Forming cover portion 22 and base portion 24 of arelatively transparent material allows light to pass through culturevessel 20 which may allow culture vessel 20 to remain in the closedposition when placed within such a growth or development room. Accordingto an exemplary embodiment, cover portion 22 and base portion 24 areformed of a plastic material such as clear polystyrene. According to thevarious alternative embodiments, cover portion 22 and base portion 24may be formed of any known or otherwise suitable transparent material(e.g., glass, etc.). According to still further alternative embodiments,one or more of cover portion 22 and base portion 24 may be formedentirely or partially of a relatively non-transparent material.

According to an exemplary embodiment, culture vessels 20 include anidentification device (not shown), such as a barcode, a radio frequencyidentification (RFID) tag or the like upon which certain informationrelating to the particular culture vessel 20 may be stored. For example,the identification device may contain information pertaining to whatstage in the production process the contents of culture vessel 20 are atso that an culture vessel 20 does not inadvertently repeat and/or miss astep in the production process. Further, the identification device maycontain information pertaining to the attributes (e.g., species, etc.)of the plant embryos being developed in culture vessel 20.

One or more receiving devices or readers (not shown) can be provided atapparatus 10 to obtain the information stored upon the identificationdevice corresponding to a particular culture vessel 20. Such a readercan be coupled to controller 90 of apparatus 10 so that informationabout the delivered culture vessels 20 can be used in determining whatoperation will be performed on culture vessels 20 at operational station14 and/or so that information about the delivered culture vessels 20 canbe displayed or be otherwise made available to an operator at apparatus10.

According to an exemplary embodiment, the identification and readerdevices may be part of a larger indexing or warehousing system. Forexample, in industrial applications it may be desirable to track thelocation and/or progress of various culture vessels 20 at any given timein the production process. Such a warehousing system may be incorporatedinto an automated transport system that selectively moves culturevessels 20 to and from various locations in the plant including a growthor development room and apparatus 10.

In operation, first rack system 40 of input station 14 is configured toreceive a first set of culture vessels 20, referred to as a first batchof culture vessels 20. As detailed above, the delivery of the firstbatch of culture vessels 20 to input station 14 may be automated ormanual. Culture vessels 20 may be empty when they arrive at inputstation 14 (e.g., if undergoing an initial base media plating operation,etc.) or may be supporting contents therein (e.g., if undergoing asubsequent inspection operation, etc.). Once arranged within first racksystem 40, culture vessels 20 are moved one-by-one by transport system18 to operational station 14 to undergo a first operation (e.g., aplating operation, etc.). After the first operation is completed atoperational station 14, the culture vessel 20 is moved to output station16 by transport system 18 and arranged in second rack system 82. Afterall of the culture vessels 20 have undergone the first operation, andare arranged at second rack system 82, the batch of culture vessels 20is moved away from apparatus 10 and taken to a growth or developmentroom. Alternatively, the batch of culture vessels 20 can be moved fromsecond rack system 82 to an auxiliary operation system (e.g., a massharvesting module, a washing module, etc.) that is coupled to apparatus10.

After a period of time (e.g., a week, six weeks, etc.), the same batchof culture vessels 20 are brought back to apparatus 10 and arrangedwithin first rack system 40. Once arranged within first rack system 40,culture vessels 20 are moved one-by-one by transport system 18 tooperational station 14 to undergo a second operation (e.g., aninspection operation, etc.). After the second operation is completed atoperational station 14, the culture vessel 20 is moved to output station16 by transport system 18 and arranged in second rack system 82. Afterall of the culture vessels 20 have undergone the second operation, andare arranged at second rack system 82, the batch of culture vessels 20is moved away from apparatus 10 and once again taken to a growth ordevelopment room. This process can be repeated until all of the steps ofthe production process have been completed.

Next, the method of preparing plant tissue for plant production will bediscussed using plant embryos as an example. For purposes of thisdisclosure, the method has been divided into seven steps. According tothe various alternative embodiments, the number of steps may be greateror less than seven. For example, additional preliminary steps may beadded relating to the development of the embryogenic tissue. Further,certain steps may be eliminated or done in combination with other steps.In step 1, a batch of empty culture vessels 20 are loaded into apparatus10 at input station 12. The culture vessels 20 are moved to operationalstation 14 at which time manipulation system 52 removes cover portion 22of culture vessel 20 from base portion 24. With cover portion 22removed, dispensing system 50 applies a base media to culture vessel 20via nozzle 72. Cover portion 22 is then returned to base portion 24 andculture vessel is moved to output station 16. Referring to FIGS.12A-12E, the dispensing operation is shown sequentially according to anexemplary embodiment. This is repeated for all of the culture vessels 20within this batch.

In step 2, the same batch of culture vessels 20 are again loaded intoapparatus 10 at input station 12. The culture vessels 20 are moved tooperational station 14 at which time manipulation system 52 (usingvacuum heads 56) removes cover portion 22 of culture vessel 20 from baseportion 24. With cover portion 22 removed, dispensing system 50 appliesa embryogenic tissue to the base media already within culture vessel 20via nozzle 72. When applying the embryogenic tissue, robotic arm 76 maymove nozzles 72 at a speed between approximately 10 millimeters persecond (mm/sec) and approximately 800 mm/sec. For example, robotic arm76 may move nozzles 72 laterally across the width of culture vessel 20at a speed of approximately 75 mm/sec.

According to an exemplary embodiment, the embryogenic tissue is “bulkedup” in a bioreactor that is coupled directly to pump 74 which is in turncoupled nozzle 72. According to an exemplary embodiment, betweenapproximately one and ten embryogenic tissue channels or lanes areformed in culture vessel 20. For example, approximately six embryogenictissue lanes may be formed in culture vessel 20 (e.g., one created byeach nozzle 72, etc.). Once the embryogenic tissue is deposited, coverportion 22 is returned to base portion 24 and culture vessel is moved tooutput station 16. This is repeated for all of the culture vessels 20within this batch. At this time, the batch of culture vessels 20 ismoved to a growth or development room where it will stay for an extendedperiod of time.

In step 3, the same batch of culture vessels 20 are again loaded intoapparatus 10 at input station 12. The culture vessels 20 are moved tooperational station 14 at which time cameras 55 of imaging system 54 areused to capture images of the plant embryos. This data is sent tocontroller 90 and is used for inspection and monitoring purposes. Theinspection of the plant embryos may be conducted with culture vessel 20in the closed position since culture vessel 20 is formed of a relativelytransparent material. Alternatively, the inspection can be conductedwith culture vessel 20 in the open position. Once the inspection iscompleted, culture vessel 20 is moved to output station 16. This isrepeated for all of the culture vessels 20 within this batch. At thistime, the batch of culture vessels 20 is moved to a growth ordevelopment room where it will stay for another extended period of time.

In step 4, the same batch of culture vessels 20 are again loaded intoapparatus 10 at input station 12. The culture vessels 20 are moved tooperational station 14 for harvesting. Apparatus 10 may provide for themass harvesting of the plant embryos or alternatively may provide forplant embryo sorting before harvesting. For mass harvesting,manipulation system 52 removes cover portion 22 from base portion 24.With cover portion 22 removed, transport system 18 delivers only baseportion 24 to a mass harvesting module that is coupled to apparatus 10.For sorting before harvesting, with cover portion 22 removed,manipulation system 52 selectively collects only those plant embryodetermined by the image processing software to be worthy of harvesting.Such a determination is based upon on or more attributes of the plantembryos (e.g., size, color, texture, etc.). Once the harvesting iscompleted, the harvested plant embryos are returned to culture vessel 20and the culture vessels are stacked at output station 16. Once theoperation is completed for all culture vessels 20, the batch is moved toa conditioning room.

In step 5, the same batch of culture vessels 20 are again loaded intoapparatus 10 at input station 12. The culture vessels 20 are moved tooperational station 14 at which time cameras 55 of imaging system 54 areused to capture images of the conditioned plant embryos. This data issent to controller 90 and is used for inspection and monitoringpurposes. The inspection of the conditioned plant embryos may beconducted with culture vessel 20 in the closed position since culturevessel 20 is formed of a relatively transparent material. Alternatively,the inspection can be conducted with culture vessel 20 in the openposition. Once the inspection is completed, culture vessel 20 is movedto output station 16. This is repeated for all of the culture vessels 20within this batch.

In step 6, the plant embryos are ready for germination. Using the dataobtained from the imaging system 54, controller 90 and the imageprocessing software are used to determine which of the plant embryos arestrong candidates for germination. Culture vessels 20 may be passedthrough operational station 14 so that manipulating system 52 can beused to collect those plant embryos selected for germination.

In step 7, another inspection operation (similar to those detailed-abovewith reference to steps 3 and 5) may take place to ensure that theselected plant embryos should be submitted for germination.

The apparatus and method detailed above provide for a relatively rapid,consistent and highly efficient production process for plant embryosthat reduces the costs associated therewith since most, if not all, ofthe steps the production process can be conducted using a singleapparatus.

In addition, because the apparatus can be partially or fully automated,human involvement during the production process is minimized. As aresult, (1) fewer humans are necessary to conduct the various operations(e.g., plating, etc.) since the apparatus and method are capable ofproducing thousands of embryos; (2) there is less chance ofcontamination caused by human contact with the embryos; (3) greaterconsistency can be achieved in the production process which leads tobetter quality control; and (4) there is better control of theproduction process since all operator input variables are handled by thecontroller.

It should be noted that according to one exemplary embodiment, theembryonic tissue is from a conifer. For example, the conifer may bepine. Further still, the pine may be a Loblolly pine.

According to other exemplary embodiments, the coniferous tree may beselected from the group consisting of Eastern white pine, Western white,Sugar pine, Red pine, Pitch pine, Jack pine, Longleaf pine, Shortleafpine, Loblolly pine, Slash pine, Virginia pine, Ponderosa pine, Jeffreypine, Pond pine, and Lodgepole pine, Radiata pine and hybrid crossesthereof. In another preferred embodiment, the coniferous tree isselected from the group consisting of, but not limited to, Abies alba,Abies amabilis, Abies balsamea, Abies bornmuelleriana, Abies concolor,Abies fraseri, Abies grandis, Abies koreana, Abies lasiocarpa, Abiesnordmanniana, Abies procera, Araucaria angustifolia, Araucaria araucana,Araucaria bidwillii, Araucaria cunninghamii, Cedrus atlantica, Cedrusdeodara, Chamaecyparis lawsoniana, Chamaecyparis pisifera, Cryptomeriajaponica, Cuppressocyparis leylandii, Larix decidua, Larix occidentalis,Metasequoia glyptostroboides, Picea abies, Picea engelmannii, Piceaglauca, Picea mariana, Picea pungens, Picea rubens, Picea sitchensis,Pinus banksiana, Pinus caribaea, Pinus contorta, Pinus echinata, Pinusedulis, Pinus elliotii, Pinus jeffreyi, Pinus korariensis, Pinuslambertiana, Pinus merkusii, Pinus monticola, Pinus nigra, Pinuspalustris, Pinus pinaster, Pinus ponderosa, Pinus rigida, Pinus radiata,Pinus resinosa, Pinus serotina, Pinus strobus, Pinus sylvestris, Pinustaeda, Pinus virginiana, Pseudotsuga menziesii, Sequoia sempervirens,Sequoiadendron giganteum, Taxodium ascends, Taxodium distichum, Taxusbaccata, Taxus brevifolia, Taxus cuspidata, Thuja occidentalis, Thujaplicata, Tsuga canadensis, Tsuga heterophylla, and hybrid crossesthereof.

Specific examples of each of such coniferous tree includes: Abies alba,European silver fir; Abies amabilis, Pacific silver fir; Abies balsamea,Balsam fir; Abies bornmuelleriana, Turkish fir; Abies concolor, Whitefir; Abies fraseri, Fraser fir; Abies grandis, Grand fir; Abies koreana,Korean fir; Abies lasiocarpa, Alpine fir; Abies nordmanniana, Nordmanfir; Abies procera, Noble fir; Araucaria angustifolia, Parana pine;Araucaria araucana, Monkeypuzzle tree; Araucaria bidwillii, Bunya pine;Araucaria cunninghamii, Hoop pine; Cedrus atlantica, Atlas cedar; Cedrusdeodara, Deodar cedar; Chamaecyparis lawsoniana, Port-Orford-cedar;Chamaecyparis pisifera, Sawara cypress; Cryptomeria japonica, Japanesecedar (Japanese cryptomeria); Cuppressocyparis leylandii, LeylandCypress; Larix decidua, European larch; Larix occidentalis, Westernlarch; Metasequoia glyptostroboides, Dawn redwood; Picea abies, Norwayspruce; Picea engelmannii, Englemann spruce; Picea glauca, White spruce;Picea mariana, Black spruce; Picea pungens, Colorado blue spruce; Picearubens, Red spruce; Picea sitchensis, Sitka spruce; Pinus banksiana,Jack pine; Pinus caribaea, Caribbean pine; Pinus contorta, lodgepolepine; Pinus echinata, Shortleaf pine; Pinus edulis, Pinyon pine; Pinuselliotii, Slash pine; Pinus jeffreyi, Jeffrey Pine; Pinus korariensis,Korean pine; Pinus lambertiana, Sugar pine; Pinus merkusii, Sumatranpine; Pinus monticola, Western white pine; Pinus nigra, Austrian pine;Pinus palustris, Longleaf pine; Pinus pinaster, Maritime pine; Pinusponderosa, Ponderosa pine; Pinus rigida, Pitch pine; Pinus radiata,Radiata pine; Pinus resinosa, Red pine; Pinus serotina, Pond pine; Pinusstrobus, Eastern white pine; Pinus sylvestris, Scots (Scotch) pine;Pinus taeda, Loblolly pine; Pinus virginiana, Virginia pine; Pseudotsugamenziesii, Douglas-fir; Sequoia sempervirens, Redwood; Sequoiadendrongiganteum, Sierra redwood; Taxodium ascends, Pond cypress; Taxodiumdistichum, Bald cypress; Taxus baccata, European yew; Taxus brevifolia,Pacific or Western yew; Taxus cuspidata, Japanese yew; Thujaoccidentalis, Northern white-cedar; Thuja plicata, Western red cedar;Tsuga canadensis, Eastern hemlock; Tsuga heterophylla, Western hemlock.

According to another exemplary embodiment, the coniferous plant tissuemay be a Southern Yellow pine. According to yet another exemplaryembodiment, the Southern Yellow pine may be selected from the groupconsisting of Pinus taeda, Pinus serotina, Pinus palustris, and Pinuselliottii.

It should further be noted that the apparatus and method detailed aboveare not limited, however, to the production of only coniferous treetissues and somatic embryos. According to another exemplary embodiment,the plant tissue, such as embryogenic tissue or a somatic embryo may befrom a tree selected from the group consisting of chestnut, ash, beech,basswood, birch, black cherry, black walnut/butternut, chinkapin,cottonwood, elm, eucalyptus, hackberry, hickory, holly, locust,magnolia, maple, oak, poplar, red alder, royal paulownia, sassafras,sweetgum, sycamore, tupelo, willow, and yellow-poplar, and intra- andinter-species hybrid crosses thereof.

It should further be noted that the plant tissue production apparatusand the methods detailed above are not limited to the development ofsomatic plant embryos, but rather may be applicable to any of variety ofplant tissue development processes. For example, the plant tissueproduction apparatus and the methods detailed above may be used withembryogenic tissues, organogenic tissue, vegetative tissue and/or seeds.Specific examples of tissue cultures include, but are not limited to,apical meristems, callus, petioles, leaf pieces, PLBs (i.e.,protocorm-like bodies), protoplasts, root tips, etc. Any such tissue,such as a tissue suspended in a liquid medium, may be distributed acrossthe culture vessel for further growth, inspection and/or datacollection. One specific example, would be to distribute vegetativepropagules of Eucalyptus ready for differentiation into plantlets acrossthe tissue culture medium surface. After a prescribed time ondifferentiation medium, successful plantlets may be identifiable via thecamera system, counted and then either transferred onto another vesselto finish growth in-vitro or be harvested for ex-vitro grow-out in agreenhouse. Another specific example, would be to use the plant tissueproduction apparatus and methods detailed above to distribute arelatively small or relatively difficult to germinate seed. For example,mutation screening in Arabidopsis could be preformed at a much largerscale enabling multiple seedlings (e.g., tens of thousands, etc.) to bescreened efficiently.

It is also important to note that the construction and arrangement ofthe elements of plant tissue production apparatus 10 as shown in theexemplary embodiments is illustrative only. Although only a fewembodiments of the present inventions have been described in detail inthis disclosure, those skilled in the art who review this disclosurewill readily appreciate that many modifications are possible (e.g.,variations in sizes, dimensions, structures, shapes and proportions ofthe various elements, values of parameters, mounting arrangements, useof materials, colors, orientations, etc.) without materially departingfrom the novel teachings and advantages of the subject matter recited.For example, elements shown as integrally formed may be constructed ofmultiple parts or elements. It should be noted that the elements and/orassemblies of the plant tissue production apparatus may be constructedfrom any of a wide variety of materials that provide sufficient strengthor durability, in any of a wide variety of colors, textures andcombinations. Accordingly, all such modifications are intended to beincluded within the scope of the present inventions. Othersubstitutions, modifications, changes and omissions may be made in thedesign, operating conditions and arrangement of the preferred and otherexemplary embodiments without departing from the spirit of the appendedclaims.

The order or sequence of any process or method steps may be varied orre-sequenced according to alternative embodiments. Anymeans-plus-function clause is intended to cover the structures describedherein as performing the recited function and not only structuralequivalents but also equivalent structures. Other substitutions,modifications, changes and omissions may be made in the design,operating configuration and arrangement of the preferred and otherexemplary embodiments without departing from the spirit of the appendedclaims.

What is claimed is:
 1. An apparatus for preparing plant tissue for plantproduction, the apparatus comprising: a first station including a firstrack system configured to support at least one culture vessel; a secondstation including an automated member configured to manipulate the atleast one culture vessel and further comprising a dispensing systemconfigured to dispense a substance to the at least one culture vesselwhen in the open position; and a third station having a second racksystem configured to support the at least one culture vessel after beingmanipulated by the automated member, wherein the automated member isconfigured to manipulate the at least one culture vessel by selectivelymoving the at least one culture vessel between an open position and aclosed position, wherein the automated member comprises a robotic armprovided at the second station and having at least one degree offreedom, wherein the robotic arm includes vacuum heads coupled to avacuum system, the vacuum heads configured to engage the at least oneculture vessel, wherein the dispensing system is configured to dispensea base media to the at least one culture vessel, wherein the dispensingsystem is further configured to dispense at least one of an embryogenictissue, organogenic tissue and vegetative tissue to the at least oneculture vessel, and wherein the at least one of the embryogenic tissue,organogenic tissue and vegetative tissue is in the form of a liquidsuspension culture and the dispensing system is in fluid communicationwith at least one reservoir containing the suspension culture.
 2. Theapparatus of claim 1, wherein the first rack system and the second racksystem are configured to support a plurality of culture vessels stackedin a substantially vertical direction, the first rack system being aninput rack system and the second rack system being an output racksystem.
 3. The apparatus of claim 2, further comprising a transportsystem configured to move the at least one culture vessel between thefirst station, the second station and the third station.
 4. Theapparatus of claim 3, wherein the first rack system includes ade-stacking system configured to present one culture vessel at a time tothe transport system for movement to the second station and the secondrack system includes a re-stacking system configured to receive the atleast one culture vessel from the second station.
 5. The apparatus ofclaim 1, further comprising a housing substantially enclosing the firststation, the second station and the third station, the housing providinga substantially sterile environment for the first station, the secondstation and the third station.
 6. The apparatus of claim 1, furthercomprising a programmable controller for controlling the movement of theautomated member, the programmable controller supporting differentprograms associated with different processes in the plant tissuedevelopment.
 7. The apparatus of claim 6, wherein the second station isselectively reconfigurable to accommodate the programs relating to thedifferent processes in the plant tissue development.
 8. The apparatus ofclaim 1, further comprising an imaging system configured to monitorplant tissue development within the at least one culture vessel.
 9. Theapparatus of claim 8, wherein the imaging system includes at least onecamera provided at the second station to capture an image of the plantdevelopment.
 10. The apparatus of claim 8, further comprising a datastorage device configured to receive data signals from the imagingsystem.
 11. The apparatus of claim 10, wherein the data storage devicereceives a first signal relating to embryo size and a second signalrelating to embryo quantity.
 12. The apparatus of claim 11, furthercomprising a processing unit that assesses the quality of the plantembryos based upon at least one predetermined parameter and at least oneof the first signal and the second signal, the at least onepredetermined parameter being at least one of a physical property and achemical property.
 13. An apparatus for preparing plant tissue containedwithin a culture vessel for plant production, the apparatus comprising:a member configured to move the culture vessel between an open positionand a closed position, the member comprising a robotic arm that includesvacuum heads coupled to a vacuum system, the vacuum heads configured toengage the culture vessel; a dispensing system configured to dispense asubstance to the culture vessel when in the open position; an imagingsystem configured to capture an image of the plant tissue; and acontroller coupled to the member, the dispensing system and the imagingsystem for automating the operation of each.
 14. The apparatus of claim13, further comprising a first rack system and a transport system, thefirst rack system being configured to support a plurality of culturevessels, the transport system being configured to transport theplurality of culture vessels to an area having at least one of themember, the dispensing system and the imaging system.
 15. The apparatusof claim 13, wherein the plant tissue is at least one of a somaticembryo, embryogenic tissue, organogenic tissue, vegetative tissue andseed.
 16. The apparatus of claim 1, wherein the vacuum heads are furtherconfigured to hold a cover of the at least one culture vessel away froma base of the at least one culture vessel.
 17. The apparatus of claim 1,wherein the robotic arm is configured to move in a substantiallyvertical direction.
 18. The apparatus of claim 1, wherein the vacuumheads are further configured to hold a cover of the culture vessel awayfrom a base of the culture vessel.
 19. The apparatus of claim 13,wherein the robotic arm is configured to move in a substantiallyvertical direction.